Diagnostic system and methods of using and manufacturing the same

Abstract

Provided are a diagnostic system and methods for detecting the presence or absence of one or more targets that represent virus infection, inflammatory diseases and/or respiratory disorders by optically and noninvasively observing changing in color of the diagnostic system upon binding of the antigen of interest. Exemplary diagnostic systems and methods include detection of SARS-CoV-2 virus S protein or receptor of advanced glycation end products (RAGE) for diagnosis of COVID-19 infection or inflammatory/respiratory diseases.

We claim: 
     
         1 . A diagnostic system, comprising:
 at least one layer of fibrous material;   a color changing moiety positioned on or within the at least one layer of fibrous material, wherein the color changing moiety is comprised of
 gold particles, 
 at least one linker linked to the gold particles, 
 an anchor protein linked to the at least one linker, and 
 at least one antibody or antibody fragment conjugated to the anchor protein, 
 wherein the at least one antibody or antibody fragment has an affinity for an antigen of interest, and 
   wherein the color changing moiety changes color when the at least one antibody or antibody fragment binds to the antigen of interest.   
     
     
         2 . The diagnostic system of  claim 1 , wherein the gold particles are gold nanoparticles. 
     
     
         3 . The diagnostic system of  claim 1 , wherein the at least one linker has a sulfhydryl moiety and a carboxylic acid moiety. 
     
     
         4 . The diagnostic system of  claim 3 , wherein the at least one linker is 11-Mercaptoundecanoic acid (MUDA) or cysteine (Cys). 
     
     
         5 . The diagnostic system of  claim 1 , wherein the anchor protein is selected from protein A, protein A ZZ domain, protein G, protein L, and fragments thereof. 
     
     
         6 . The diagnostic system of  claim 1 , wherein the at least one layer of fibrous material is nonwoven. 
     
     
         7 . The diagnostic system of  claim 1 , wherein the at least one layer of fibrous material is woven. 
     
     
         8 . The diagnostic system of  claim 1 , wherein the antigen of interest is a protein having a molecular weight of about 45 to 250 kDa. 
     
     
         9 . The diagnostic system of  claim 1 , wherein the antigen of interest is SARS-CoV-2 virus S protein. 
     
     
         10 . The diagnostic system of  claim 1 , wherein the antigen of interest is receptor of advanced glycation end products (RAGE). 
     
     
         11 . The diagnostic system of  claim 1 , wherein the gold particles are immobilized on or bound to one or more fibers of the at least one layer of fibrous material. 
     
     
         12 . The diagnostic system of  claim 1 , wherein the diagnostic system is configured as a face mask wearable by a human or non-human animal subject. 
     
     
         13 . The diagnostic system of  claim 1 , wherein the diagnostic system is configured as a garment. 
     
     
         14 . The diagnostic system of  claim 1 , wherein the diagnostic system is configured as a bedding cover. 
     
     
         15 . The diagnostic system of  claim 1 , wherein the diagnostic system is configured as a wipe. 
     
     
         16 . The diagnostic system of  claim 15 , wherein the wipe is configured for use on animal skin. 
     
     
         17 . The diagnostic system of  claim 15 , wherein the wipe is configured for use on substrates other than animal skin. 
     
     
         18 . The diagnostic system of  claim 1 , wherein the diagnostic system is configured as a surface cover. 
     
     
         19 . A method for detecting one or more antigens, comprising:
 bringing a fluid or surface which is contaminated with the one or more antigens into contact with the diagnostic system of  claim 1 ; and   detecting a change in color on at least one portion of the diagnostic system when the one or more antigens become bound to the at least one antibody or antibody fragment capable of recognizing the one or more antigens.   
     
     
         20 . The method of  claim 19 , wherein the detection is made by optical observation. 
     
     
         21 . The method of  claim 19 , wherein the optical observation occurs within about 10-30 minutes of binding of the one or more antigens to the at least one antibody or antibody fragment capable of recognizing the one or more antigens. 
     
     
         22 . The method of  claim 19 , further comprising a step of assessing an amount of coupling between the antigens to antibody or antibody fragment capable of recognizing the one or more antigens. 
     
     
         23 . The method of  claim 22 , wherein the amount of coupling is calculated by comparing a degree of colorimetric change after contact with the fluid or surface with a known degree of colorimetric change after contact with a known amount of an antigen of interest. 
     
     
         24 . The method of  claim 19 , wherein the antigen of interest is a protein having a molecular weight of about 45 to 250 kDa. 
     
     
         25 . The method of  claim 19 , wherein the one or more antigens comprise SARS-CoV-2 virus S protein. 
     
     
         26 . The method of  claim 19 , wherein the one or more antigens comprise receptor of advanced glycation end products (RAGE). 
     
     
         27 . The method of  claim 19 , wherein the fluid or surface brought into contact with the diagnostic system is or comprises breath exhaled from a human or non-human animal subject. 
     
     
         28 . The method of  claim 19 , wherein the fluid or surface is or comprises a bodily discharge of a subject. 
     
     
         29 . The method of  claim 19 , wherein the fluid of interest does not travel across or through a surface of the at least one layer of fibrous material. 
     
     
         30 . A method of manufacturing the diagnostic system of  claim 1 , comprising the steps of:
 synthesizing gold particles (Au Ps) on at least one piece of fabric by
 adding the at least one piece of fabric to a HAuCl 4  solution to nucleate Au 0  directly on the fabric to form Au Ps fabric, 
 conjugating at least one linker to the Au Ps fabric to form linker-coupled gold particles fabric; 
 linking at least one anchoring protein to the linker; and 
 conjugating at least one antibody or antibody fragment capable of recognizing the one or more antigens to the at least one anchoring protein. 
   
     
     
         31 . The method of  claim 30 , wherein the gold particles are gold nanoparticles. 
     
     
         32 . The method of  claim 30 , wherein the at least one linker has a sulfhydryl moiety and a carboxylic acid moiety. 
     
     
         33 . The method of  claim 32 , wherein the at least one linker is 11-Mercaptoundecanoic acid (MUDA) or cysteine (Cys). 
     
     
         34 . The method of  claim 30 , wherein the at least one piece of fabric is nonwoven. 
     
     
         35 . The method of  claim 30 , wherein the at least one piece of fabric is woven. 
     
     
         36 . The method of  claim 30 , wherein the HAuCl 4  solution comprises a reducing agent. 
     
     
         37 . The method of  claim 30 , further comprising pretreating the at least one piece of fabric in a solution containing a nucleating agent. 
     
     
         38 . The method of  claim 37 , wherein the nucleating agent is ZnO nanoparticles (ZnO NPs). 
     
     
         39 . The method of  claim 30 , wherein the at least one anchoring protein is selected from protein A, protein A ZZ domain, protein G, protein L, and fragments thereof. 
     
     
         40 . The method of  claim 30 , further comprising adding the Au Ps fabric to a HAuCl 4  solution to further grow nucleated Au Ps.